title: Profiling humoral immune responses to P. falciparum infection with protein microarrays.
abstract: A complete description of the serological response following exposure of humans to complex pathogens is lacking and approaches suitable for accomplishing this are limited. Here we report, using malaria as a model, a method which elucidates the profile of antibodies that develop after natural or experimental infection or after vaccination with attenuated organisms, and which identifies immunoreactive antigens of interest for vaccine development or other applications. Expression vectors encoding 250 Plasmodium falciparum (Pf) proteins were generated by PCR/recombination cloning; the proteins were individually expressed with >90% efficiency in Escherichia coli cell-free in vitro transcription and translation reactions, and printed directly without purification onto microarray slides. The protein microarrays were probed with human sera from one of four groups which differed in immune status: sterile immunity or no immunity against experimental challenge following vaccination with radiation-attenuated Pf sporozoites, partial immunity acquired by natural exposure, and no previous exposure to Pf. Overall, 72 highly reactive Pf antigens were identified. Proteomic features associated with immunoreactivity were identified. Importantly, antibody profiles were distinct for each donor group. Information obtained from such analyses will facilitate identifying antigens for vaccine development, dissecting the molecular basis of immunity, monitoring the outcome of whole-organism vaccine trials, and identifying immune correlates of protection.
authors.affiliation: Naval Medical Research Center, Silver Spring, MD, USA.
authors.forename: Denise L
authors.initials: DL
authors.lastname: Doolan
authors.forename: Yunxiang
authors.initials: Y
authors.lastname: Mu
authors.forename: Berkay
authors.initials: B
authors.lastname: Unal
authors.forename: Suman
authors.initials: S
authors.lastname: Sundaresh
authors.forename: Siddiqua
authors.initials: S
authors.lastname: Hirst
authors.forename: Conrad
authors.initials: C
authors.lastname: Valdez
authors.forename: Arlo
authors.initials: A
authors.lastname: Randall
authors.forename: Douglas
authors.initials: D
authors.lastname: Molina
authors.forename: Xiaowu
authors.initials: X
authors.lastname: Liang
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Freilich
authors.forename: J Aggrey
authors.initials: JA
authors.lastname: Oloo
authors.forename: Peter L
authors.initials: PL
authors.lastname: Blair
authors.forename: Joao C
authors.initials: JC
authors.lastname: Aguiar
authors.forename: Pierre
authors.initials: P
authors.lastname: Baldi
authors.forename: D Huw
authors.initials: DH
authors.lastname: Davies
authors.forename: Philip L
authors.initials: PL
authors.lastname: Felgner
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000276:immunology*; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology; D030562:Databases, Protein; D006801:Humans; D017780:Malaria Vaccines / Q000276:immunology; D016778:Malaria, Falciparum / Q000235:genetics / Q000276:immunology*; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D017780:Malaria Vaccines; D015800:Protozoan Proteins